title: Purification, crystallization and preliminary crystallographic studies of C-terminal RNA recognition motif (RRM-3) of human ELAV-type RNA-binding protein 3 (ETR-3).
abstract: Human embryonically lethal abnormal vision (ELAV)-type RNA-binding protein 3 (ETR-3) has been implicated in many aspects of RNA-processing events including alternative splicing, stability, editing and translation. RNA recognition motif 3 (RRM-3) is an independent C-terminal RNA-binding domain of ETR-3 that preferentially binds to UG-rich repeats of the nuclear or cytoplasmic pre-mRNA, and along with the other domains mediates the inclusion of cardiac troponin T (c-TNT) exon 5 in embryonic muscle, which is otherwise excluded in the adult. In the present study, RRM-3 was cloned, overexpressed, purified and crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to 3 Å resolution at the home source and belonged to space group P2₁3, with unit-cell parameters a=b=c=118.5 Å, α=β=γ=90°. There were two molecules of RRM-3 in the asymmetric unit and the calculated Matthews coefficient (VM) was 6.35 Å3 Da(-1), with a solvent content of 80.62%. Initial phases were determined by molecular replacement.
authors.affiliation: Structural and Computational Biology Group, International Center for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi 110 067, India.
authors.forename: Maruthi
authors.initials: M
authors.lastname: Kashyap
authors.forename: Ashwani
authors.initials: A
authors.lastname: Sharma
authors.forename: Neel Sarovar
authors.initials: NS
authors.lastname: Bhavesh
mesh_terms: D020816:Amino Acid Motifs; D000067878:CELF Proteins; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000067878:CELF Proteins; C117363:CELF2 protein, human; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins
keywords: C-terminal RNA recognition motif; ETR-3; RRM-3; human ELAV-type RNA-binding protein 3